Celery cultivar ADS-19

ABSTRACT

A celery cultivar, designated ADS-19, is disclosed. The invention relates to the seeds of celery cultivar ADS-19, to the plants of celery cultivar ADS-19 and to methods for producing a celery plant by crossing the cultivar ADS-19 with itself or another celery cultivar. The invention further relates to methods for producing a celery plant containing in its genetic material one or more transgenes and to the transgenic celery plants and plant parts produced by those methods. This invention also relates to celery cultivars or breeding cultivars and plant parts derived from celery cultivar ADS-19, to methods for producing other celery cultivars, lines or plant parts derived from celery cultivar ADS-19 and to the celery plants, varieties, and their parts derived from the use of those methods. The invention further relates to hybrid celery seeds, plants, and plant parts produced by crossing cultivar ADS-19 with another celery cultivar.

BACKGROUND OF THE INVENTION

The present invention relates to a new and distinctive celery (Apiumgraveolens var. dulce) variety, designated ADS-19. All publicationscited in this application are herein incorporated by reference.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm. These important traits may include improved flavor, increasedstalk size and weight, higher seed yield, improved color, resistance todiseases and insects, tolerance to drought and heat, and betteragronomic quality.

Practically speaking, all cultivated forms of celery belong to thespecies Apium graveolens var. dulce that is grown for its edible stalk.As a crop, celery is grown commercially wherever environmentalconditions permit the production of an economically viable yield. In theUnited States, the principal growing regions are California, Florida,Texas and Michigan. Fresh celery is available in the United Statesyear-round although the greatest supply is from November throughJanuary. For planting purposes, the celery season is typically dividedinto two seasons, summer and winter, with Florida, Texas and thesouthern California areas harvesting from November to July, and Michiganand northern California harvesting from July to October. Celery isconsumed as fresh, raw product and occasionally as a cooked vegetable.

Celery is a cool-season biennial that grows best from 60° to 65° F. (16°to 18° C.), but will tolerate temperatures from 45° to 75° F. (7° to 24°C.). Freezing damages mature celery by splitting the petioles or causingthe skin to peel, making the stalks unmarketable. This is an occasionalproblem in plantings in the winter regions, however, celery can tolerateminor freezes early in the crop.

The two main growing regions for celery in California are located alongthe Pacific Ocean: the central coast or summer production area(Monterey, San Benito, Santa Cruz and San Luis Obispo Counties) and thesouth coast or winter production area (Ventura and Santa BarbaraCounties). A minor region (winter) is located in the southern deserts(Riverside and Imperial Counties).

In the south coast, celery is transplanted from early August to Aprilfor harvest from November to mid-July; in the Santa Maria area, celeryis transplanted from January to August for harvest from April throughDecember. In the central coast, fields are transplanted from March toSeptember for harvest from late June to late December. In the southerndeserts, fields are transplanted in late August for harvest in January.

Commonly used celery varieties for coastal production include Tall Utah52-75, Conquistador and Sonora. Some shippers use their own proprietaryvarieties. Celery seed is very small and difficult to germinate. Allcommercial celery is planted as greenhouse-grown transplants. Celerygrown from transplants is more uniform than from seed and takes lesstime to grow the crop in the field. Transplanted celery is placed indouble rows on 40-inch (100-cm) beds with plants spaced between 6.0 and7 inches apart.

Celery requires a relatively long and cool growing season (Thephysiology of vegetable crops by Pressman, CAB Intl., New York, 1997).Earlier transplanting results in a longer growing season, increasedyields, and better prices. However, celery scheduled for Spring harvestoften involves production in the coolest weather conditions of Winter, aperiod during which vernalization can occur. If adequate vernalizationoccurs for the variety, bolting may be initiated. Bolting is thepremature rapid elongation of the main celery stem into a floral axis(i.e., during the first year for this normally biennial species). Ifbolting occurs, growth slows and initiation of flower stalks, or theseed stems, occurs prematurely as the plant approaches marketable size,leaving a stalk with no commercial value. Different varieties havedifferent vernalization requirements, but in the presence of bolting,the length of the seed stem can be used as a means of measuring orcomparing different varieties for the amount of bolting tolerance thatexists in each variety. The most susceptible varieties reach theirvernalization requirement earlier and have time to develop the longestseed stems, while the moderately tolerant varieties take longer to reachtheir vernalization requirement and have less time to develop a seedstem which would therefore be shorter. Under normal productionconditions, the most tolerant varieties may not achieve theirvernalization requirement and therefore not produce a measurable seedstem.

The coldest months when celery is grown in the United States areDecember, January and February. If celery is going to reach itsvernalization requirements to cause bolting, it is generally youngercelery that is exposed to this cold weather window. This celerygenerally matures in the months of April and May which constitutes whatthe celery industry calls the bolting or seeder window. The bolting orseeder window is a period where seed stems are generally going to impactthe quality of the marketable celery and this is most consistentlyexperienced in celery grown in the Southern California region. Thepresence of seed stems in celery can be considered a major marketabledefect as set forth in the USDA grade standards. If the seed stem islonger than twice the diameter of the celery stalk or eight inches, thecelery no longer meets the standards of US Grade #1. If the seed stem islonger than three times the diameter of the celery stalk, the celery isno longer marketable as US Grade #2 (United States Standards for Gradesof Celery, United States Department of Agriculture, reprinted January1997).

Celery is an allogamous biennial crop. The celery genome consists of 11chromosomes. Its high degree of out-crossing is accomplished by insectsand wind pollination. Pollinators visiting celery flowers include alarge number of wasp, bee and fly species. Celery is subject toinbreeding depression which appears to be genotype dependent, since somelines are able to withstand continuous selfing for three or fourgenerations. Crossing of inbreds results in heterotic hybrids that arevigorous and taller than sib-mated or inbred lines.

Celery flowers are protandrous, with pollen being released 3-6 daysbefore stigma receptivity. At the time of stigma receptivity the stamenswill have fallen and the two stigmata unfolded in an upright position.The degree of protandry varies, which makes it difficult to performreliable hybridization, due to the possibility of accidental selfing.

Celery flowers are very small, significantly precluding easy removal ofindividual anthers. Furthermore, different developmental stages of theflowers in umbels makes it difficult to avoid uncontrolled pollinations.The standard hybridization technique in celery consists of selectingflower buds of the same size and eliminating the older and youngerflowers. Then, the umbellets are covered with glycine paper bags for a5-10 day period, during which the stigmas become receptive. At the timethe flowers are receptive, available pollen or umbellets shedding pollenfrom selected male parents are rubbed on to the stigmas of the femaleparent.

Celery plants require a period of vernalization while in the vegetativephase in order to induce seed stalk development. A period of 6-10 weeksat 5-8° C. is usually adequate. However, unless plants are beyond ajuvenile state or a minimum of 4 weeks old they may not be receptive tovernalization. Due to a wide range of responses to the cold treatment,it is often difficult to synchronize crossing, since plants will flowerat different times. However, pollen can be stored for 6-8 months at−10EC in the presence of silica gel or calcium chloride with a viabilitydecline of only 20-40%, thus providing flexibility to perform crossesover a longer time.

For selfing, the plant or selected umbels are caged in cloth bags. Theseare shaken several times during the day to promote pollen release.Houseflies (Musca domestica) can also be introduced weekly into the bagsto perform pollinations.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, and recurrent selection.

The complexity of inheritance influences choice of the breeding method.Backcross breeding is used to transfer one or a few favorable genes fora highly heritable trait into a desirable cultivar. This approach hasbeen used extensively for breeding disease-resistant cultivars. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes. The use of recurrentselection in self-pollinating crops depends on the ease of pollination,the frequency of successful hybrids from each pollination, and thenumber of hybrid offspring from each successful cross.

Each breeding program should include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard, theoverall value of the advanced breeding lines, and the number ofsuccessful cultivars produced per unit of input (e.g., per year, perdollar expended, etc.).

Promising advanced breeding lines are thoroughly tested and compared toappropriate standards in environments representative of the commercialtarget area(s) for three years at least. The best lines are candidatesfor new commercial cultivars; those still deficient in a few traits areused as parents to produce new populations for further selection.

These processes, which lead to the final step of marketing anddistribution, usually take from ten to twenty years from the time thefirst cross or selection is made. Therefore, development of newcultivars is a time-consuming process that requires precise forwardplanning, efficient use of resources, and a minimum of changes indirection.

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

The goal of celery plant breeding is to develop new, unique and superiorcelery cultivars. The breeder initially selects and crosses two or moreparental lines, followed by repeated selfing and selection, producingmany new genetic combinations. The breeder can theoretically generatebillions of different genetic combinations via crossing, selfing andmutations. The breeder has no direct control at the cellular level.Therefore, two breeders will never develop the same line, or even verysimilar lines, having the same celery traits.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions, and further selections arethen made, during and at the end of the growing season. The cultivarsthat are developed are unpredictable. This unpredictability is becausethe breeder's selection occurs in unique environments, with no controlat the DNA level (using conventional breeding procedures), and withmillions of different possible genetic combinations being generated. Abreeder of ordinary skill in the art cannot predict the final resultinglines he develops, except possibly in a very gross and general fashion.The same breeder cannot produce the same line twice by using the exactsame original parents and the same selection techniques. Thisunpredictability results in the expenditure of large research monies todevelop superior celery cultivars.

The development of commercial celery cultivars requires the developmentof celery varieties, the crossing of these varieties, and the evaluationof the crosses. Pedigree breeding and recurrent selection breedingmethods are used to develop cultivars from breeding populations.Breeding programs combine desirable traits from two or more varieties orvarious broad-based sources into breeding pools from which cultivars aredeveloped by selfing and selection of desired phenotypes. The newcultivars are crossed with other varieties and the hybrids from thesecrosses are evaluated to determine which have commercial potential.

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents which possess favorable, complementary traits, are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁'s or by intercrossing two F₁'s (sib mating). Selection of the bestindividuals is usually begun in the F₂ population; then, beginning inthe F₃, the best individuals in the best families are selected.Replicated testing of families, or hybrid combinations involvingindividuals of these families, often follows in the F₄ generation toimprove the effectiveness of selection for traits with low heritability.At an advanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease as new cultivars.

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued.

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror line that is the recurrent parent. The source of the trait to betransferred is called the donor parent. The resulting plant is expectedto have the attributes of the recurrent parent (e.g., cultivar) and thedesirable trait transferred from the donor parent. After the initialcross, individuals possessing the phenotype of the donor parent areselected and repeatedly crossed (backcrossed) to the recurrent parent.The resulting plant is expected to have the attributes of the recurrentparent (e.g., cultivar) and the desirable trait transferred from thedonor parent.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F₂ to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F₂ individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F₂ plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. There are many laboratory-based techniques available forthe analysis, comparison and characterization of plant genotype; amongthese are Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length polymorphisms (AFLPs), Simple Sequence Repeats(SSRs—which are also referred to as Microsatellites), and SingleNucleotide Polymorphisms (SNPs).

Isozyme Electrophoresis and RFLPs have been widely used to determinegenetic composition. Shoemaker and Olsen, (Molecular Linkage Map ofSoybean (Glycine max) p 6.131-6.138 in S. J. O'Brien (ed) Genetic Maps:Locus Maps of Complex Genomes, Cold Spring Harbor Laboratory Press, ColdSpring Harbor, N.Y., (1993)) developed a molecular genetic linkage mapthat consisted of 25 linkage groups with about 365 RFLP, 11 RAPD, threeclassical markers and four isozyme loci. See also, Shoemaker, R. C.,RFLP Map of Soybean, p 299-309, in Phillips, R. L. and Vasil, I. K.,eds. DNA-Based Markers in Plants, Kluwer Academic Press, Dordrecht, theNetherlands (1994).

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. Forexample, Diwan and Cregan described a highly polymorphic microsatellitelocus in soybean with as many as 26 alleles. (Diwan, N. and Cregan, P.B., Theor. Appl. Genet. 95:22-225,1997.) SNPs may also be used toidentify the unique genetic composition of the invention and progenyvarieties retaining that unique genetic composition. Various molecularmarker techniques may be used in combination to enhance overallresolution.

Molecular markers, which include markers identified through the use oftechniques such as Isozyme Electrophoresis, RFLPs, RAPDs, AP-PCR, DAF,SCARs, AFLPs, SSRs, and SNPs, may be used in plant breeding. One use ofmolecular markers is Quantitative Trait Loci (QTL) mapping. QTL mappingis the use of markers which are known to be closely linked to allelesthat have measurable effects on a quantitative trait. Selection in thebreeding process is based upon the accumulation of markers linked to thepositive effecting alleles and/or the elimination of the markers linkedto the negative effecting alleles from the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select toward the genome of the recurrent parent and against themarkers of the donor parent. This procedure attempts to minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called genetic marker enhancedselection or marker-assisted selection. Molecular markers may also beused to identify and exclude certain sources of germplasm as parentalvarieties or ancestors of a plant by providing a means of trackinggenetic profiles through crosses.

Mutation breeding is another method of introducing new traits intocelery varieties. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in Principles of Cultivar Development by Fehr, MacmillanPublishing Company, 1993.

The production of double haploids can also be used for the developmentof homozygous varieties in a breeding program. Double haploids areproduced by the doubling of a set of chromosomes from a heterozygousplant to produce a completely homozygous individual. For example, seeWan et al., Theor. Appl. Genet, 77:889-892, 1989.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., Principles of Plant Breeding John Wiley and Son, pp.115-161, 1960; Allard, 1960; Simmonds, 1979; Sneep et al., 1979; Fehr,1987; ACarrots and Related Vegetable Umbelliferae@, Rubatzky, V. E., etal., 1999).

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

Celery in general is an important and valuable vegetable crop. Thus, acontinuing goal of celery plant breeders is to develop stable, highyielding celery cultivars that are agronomically sound. The reasons forthis goal are obviously to maximize the amount of yield produced on theland. To accomplish this goal, the celery breeder must select anddevelop celery plants that have the traits that result in superiorcultivars.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools, and methods which are meant to beexemplary and illustrative, not limiting in scope. In variousembodiments, one or more of the above-described problems have beenreduced or eliminated, while other embodiments are directed to otherimprovements.

According to the invention, there is provided a novel celery cultivar,designated ADS-19. This invention thus relates to the seeds of celerycultivar ADS-19, to the plants of celery cultivar ADS-19 and to methodsfor producing a celery plant produced by crossing the celery ADS-19 withitself or another celery plant, to methods for producing a celery plantcontaining in its genetic material one or more transgenes and to thetransgenic celery plants produced by that method. This invention alsorelates to methods for producing other celery cultivars derived fromcelery cultivar ADS-19 and to the celery cultivar derived by the use ofthose methods. This invention further relates to hybrid celery seeds andplants produced by crossing celery cultivar ADS-19 with another celeryline.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of celery cultivar ADS-19. The tissue culture willpreferably be capable of regenerating plants having essentially all ofthe physiological and morphological characteristics of the foregoingcelery plant, and of regenerating plants having substantially the samegenotype as the foregoing celery plant. Preferably, the regenerablecells in such tissue cultures will be callus, protoplasts, meristematiccells, leaves, pollen, embryos, roots, root tips, anthers, pistils,flowers, seeds, petioles and suckers. Still further, the presentinvention provides celery plants regenerated from the tissue cultures ofthe invention.

Another aspect of the invention is to provide methods for producingother celery plants derived from celery cultivar ADS-19. Celerycultivars derived by the use of those methods are also part of theinvention.

The invention also relates to methods for producing a celery plantcontaining in its genetic material one or more transgenes and to thetransgenic celery plant produced by those methods.

In another aspect, the present invention provides for single geneconverted plants of ADS-19. The single transferred gene may preferablybe a dominant or recessive allele. Preferably, the single transferredgene will confer such traits as male sterility, herbicide resistance,insect resistance, modified fatty acid metabolism, modified carbohydratemetabolism, resistance for bacterial, fungal, or viral disease, malefertility, enhanced nutritional quality and industrial usage or thetransferred gene will have no apparent value except for the purpose ofbeing a marker for variety identification. The single gene may be anaturally occurring celery gene or a transgene introduced throughgenetic engineering techniques.

The invention further provides methods for developing celery plant in acelery plant breeding program using plant breeding techniques includingrecurrent selection, backcrossing, pedigree breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection and transformation. Seeds, celery plants, and parts thereof,produced by such breeding methods are also part of the invention.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference by thestudy of the following descriptions.

DEFINITIONS

In the description and tables which follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele. An allele is any of one or more alternative form of a gene, allof which relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing. Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Blackheart. Blackheart is due to a lack of movement of sufficientcalcium that causes the plant to turn brown and begin to decay at thegrowing point of the plant. Celery in certain conditions, such as warmweather, grows very rapidly and is incapable of moving sufficientamounts of calcium to the growing point.

Bolting. The premature development of a flowering or seed stalk, andsubsequent seed, before a plant produces a food crop. Bolting istypically caused by late planting when temperatures are low enough tocause vernalization of the plants.

Bolting Period. Also known as the bolting or seeder window, andgenerally occurs in celery that is transplanted from the middle ofDecember through January and matures in April to May. The intensity andactual weeks that bolting may be observed vary from year to year, but itis generally observed in this window.

Bolting Tolerance. The amount of vernalization that is required fordifferent celery varieties to bolt is genetically controlled. Varietieswith increased tolerance to bolting require greater periods ofvernalization in order to initiate bolting. A comparison of boltingtolerance between varieties can be measured by the length of theflowering or seed stem under similar vernalization conditions.

Brown Stem. A disease caused by the bacterium Pseudomonas cichorii thatcauses petiole necrosis. Brown Stem is characterized by a firm, browndiscoloration throughout the petiole.

Celeriac or Root celery (Apium graveolens L. var. rapaceum). Means aplant that is related to celery but instead of having a thickened andsucculent leaf petiole as in celery, celeriac has an enlarged hypocotyland upper root that is the edible product.

Celery Heart. Means the center most interior petioles and leaves of thecelery stalk. They are not only the smallest petioles in the stalk, butthe youngest as well. Some varieties are considered heartless becausethey go right from very large petioles to only a couple of very smallpetioles. The heart is comprised of the petioles that are closest to themeristem of the celery stalk. Most straw and process type varieties havevery little heart development.

Consumable. Means material that is edible by humans.

Crackstem. The petiole can crack or split horizontally orlongitudinally. Numerous cracks in several locations along the petioleare often an indication that the variety has insufficient boronnutrition. A variety's ability to utilize boron is a physiologicalcharacteristic which is genetically controlled.

Dry weight .Means the weight of the celery after all water has beenremoved from celery.

Dry weight percentage. Means the calculation of the dry weight of thecelery divided by the original weight of the celery before the removalof the water.

Durable. Means a long-lasting, sturdy and resilient celery, that is ableto resist breakage and ruptures through normal harvesting, processing,packaging, shipment and usage.

Edible celery (Apium graveolens L. var. dulce). Means an Apiumgraveolens L. var. dulce celery that is considered suitable foringestion by humans based upon the flavor and the texture of the celery.

Essentially all the physiological and morphological characteristics. Aplant having essentially all the physiological and morphologicalcharacteristics means a plant having the physiological and morphologicalcharacteristics of the recurrent parent, except for the characteristicsderived from the converted gene.

External diameter. Means the average diameter of the petiole cylindermeasured from the outside of the cylinder wall to the outside of theopposite cylinder wall.

Feather Leaf. Feather Leaf is a yellowing of the lower leaves andgenerally occurs in the outer petioles but can also be found on innerpetioles of the stalk. These yellowing leaves which would normallyremain in the harvested stalk are considered unacceptable. Thesepetioles then have to be stripped off in order to meet market gradewhich effectively decreases the stalk size and yield.

Flare. The lower, generally wider portion of the petiole which isusually paler green or white.

Fusarium Yellows. A fungal soilborne disease caused by Fusariumoxysporum f. sp. apii Race 2. Infected plants turn yellow and arestunted. Some of the large roots may have a dark brown, water-soakedappearance. The water-conducting tissue (xylem) in the stem, crown, androot show a characteristic orange-brown discoloration. In the laterstages of infection, plants remain severely stunted and yellowed and maycollapse.

Gene. As used herein, “gene” refers to a segment of nucleic acid. A genecan be introduced into a genome of a species, whether from a differentspecies or from the same species, using transformation or variousbreeding techniques.

Gross Yield (Pounds/Acre). The total yield in pounds/acre of trimmedcelery plants (stalks).

Hollow tube or hollow petiole. Means the shape of the celery petiolewherein the petiole is cylindrical, nearly cylindrical, hemispherical ornearly hemispherical and hollow in the center.

Internal diameter. Means the average diameter of the petiole cylindermeasured from the inside of the cylinder wall to the inside of theopposite cylinder wall.

Leaf Celery (Apium graveolens L. var. secalinum). Means a plant that hasbeen developed primarily for leaf and seed production. Often grown inMediterranean climates, leaf celery more closely resembles celery's wildancestors. The stems are small and fragile and vary from solid to hollowand the leaves are fairly small and are generally bitter. This type isoften used for its medicinal properties and spice.

Leaf Margin Chlorosis. A magnesium deficiency producing an interveinalchlorosis which starts at the margin of leaves.

Maturity Date. Maturity in celery can be dictated by two conditions. Thefirst, or true maturity, is the point in time when the celery reachesmaximum size distribution, but before defects such as pith, yellowing,Feather Leaf or Brown Stem appear. The second, or market maturity is anartificial maturity dictated by market conditions, i.e, the marketrequirement may be for 3 dozen sizes so the field is harvested atslightly below maximum yield potential because the smaller sizes arewhat the customers prefer at that moment.

Mean straw width range. Means the range created by the comparison of themean of the narrowest straw widths as measured in several stalks ofhollow celery that can be used as a straw versus the of mean widestwidths measured in several stalks of hollow celery sticks that can beused as a straw.

MUN. MUN refers to the MUNSELL Color Chart which publishes an officialcolor chart for plant tissues according to a defined numbering system.The chart may be purchased from the Macbeth Division of KollmorgenInstruments Corporation, 617 Little Britain Road, New Windsor, N.Y.12553-6148.

Packaged. Means the celery limbs are packaged according to length andmay be packaged in any number of methods according to the specificationsof the customer. The product may be packaged and sealed in flexiblefilms, including sleeves or bags that may or may not be resealable,rigid plastic containers like clam shells, solid fiber containers, polysleeves, plastic sleeves, poly bags, plastic bags, natural decomposablebags, natural decomposable sleeves, or any combination thereof.Variations in the packaging may include different gas exchange rateswhich may occur due to different permeability or transmission propertiesof the package materials themselves or due to vents or specialized poresbuilt into the packaging. The package weight ranges from 0.25, 0.26,0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38,0.39, 0.40, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.50,0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.60, 0.61, 0.62,0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.70, 0.71, 0.72, 0.73, 0.74,0.75, 0.76, 0.77, 0.78, 0.79, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86,087, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98,0.99, 1.00, 1.01, 1.02, 1.03, 1.04, 1.05, 1.06, 1.07, 1.08, 1.09, 1.10,1.11, 1.12, 1.13, 1.14, 1.15, 1.16, 1.17, 1.18, 1.19, 1.20, 1.21, 1.22,1.23, 1.24, 1.25, 1.26, 1.27, 1.28, 1.29, 1.30, 1.31, 1.32, 1.33, 1.34,1.35, 1.36, 1.37, 1.38, 1.39, 1.40, 1.41, 1.42, 1.43, 1.44, 1.45, 1.46,1.47, 1.48, 1.49, 1.50, 1.51, 1.52, 1.53, 1.54, 1.55, 1.56, 1.57, 1.58,1.59, to 1.60 pounds or higher and including all integers and fractionsthereof.

Petiole. A petiole is the stem or limb of a leaf, the primary portion ofthe celery consumed.

Petiole depth. Means the average measurement in millimeters of the depthof the celery petiole at its narrowest point. The petiole depthmeasurement is taken from the outside of the petiole (which is the partof the petiole that faces the outside of the stalk) and is measured tothe inside of the petiole or cup or the inner most point of the petiolethat faces the center of the stalk or heart.

Inside Petiole Cup tissue or tissue enclosure Petiole Cup. Means thetissue on the inside cup of the petiole that encloses the edges of thepetiole cup and creates the hollow celery.

Petiole width. Means the average measurement of the width of the celerypetiole in millimeters at its widest point. The measurement is takenfrom the side or edge of petiole to the opposite side or edge of thepetiole. The measurement is taken 90 degrees from petiole depth.

Phthalides. Means one of the chemical compounds that is responsible forthe characteristic flavor and aroma of celery.

Plant Height. Means the height of the plant from the bottom of the baseor butt of the celery plant to the top of the tallest leaf.

Quantitative Trait Loci. Quantitative Trait Loci (QTL) refers to geneticloci that control to some degree, numerically representable traits thatare usually continuously distributed.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Ribbing. The texture of the surface of the celery petiole can vary fromsmooth to ribby depending on the variety. Ribbing is the presence ofnumerous ridges that run vertically along the petioles of the celeryplant.

Sanitized. Means washed, cleansed or sterilized hollow celery so thelimb's surface is free of dirt, insects, microbial infestation,bacterial infestation, fungal infestation or other surface contaminates.The process of sanitization involves washing the limbs in order toremove surface contamination such as dirt and insects and theutilization of a sanitization material or process in order to remove orkill surface contamination by microbial, bacterial and fungal agents.

Sanitization Treatment. Means treating the celery with a chemical orprocess so as to sanitize the celery. The chemical or process isselected from the group consisting of ascorbic acid, peroxyacetic acidalso known as TSUNAMI, sodium hypochlorite (chlorine), bromine products(sodium hypobromine), chlorine dioxide, ozone based systems, hydrogenperoxide products, trisodium phosphate, quaternary ammonium products,ultraviolet light systems, irradiation, steam, ultra heat treatments,and high pressure pasteurization.

Shear strength or pressure. Means the force in grams that a celery canwithstand prior rupturing or cutting of the wall of the celery petiole.

Seed Stem. A seed stem is the result of the elongation of the main stemof the celery, which is usually very compressed to almost non-existent,to form the flowering axis. The seed stem or flowering axis can reachseveral feet in height during full flower. The length of the seed stemis measured as the distance from the top of the basal plate (the base ofthe seed stem) to its terminus (the terminal growing point).

Single gene converted. Single gene converted or conversion plant refersto plants which are developed by a plant breeding technique calledbackcrossing or via genetic engineering wherein essentially all of thedesired morphological and physiological characteristics of a line arerecovered in addition to the single gene transferred into the line viathe backcrossing technique or via genetic engineering.

Stalk. A stalk is a single celery plant that is trimmed with the top orfoliage and the roots removed.

Straw celery. Means an Apium graveolens L. var. dulce hollow celerystick that is capable of being used to suck up a liquid.

Stringiness. Stringiness is a physiological characteristic that isgenerally associated with strings that get stuck between the consumer'steeth. There are generally two sources of strings in celery. One is thevascular bundle which can be fairly elastic and behave as a string. Thesecond is a strip of particularly strong epidermis which is located onthe surface of the ridges of the celery varieties that have ribs.

Stuffed celery. Means an Apium graveolens L. var. dulce hollow celerystick that is capable of being filled with edible products.

Suckers. Suckers are auxiliary shoots that form at the base of the stalkor within the auxiliary buds between each petiole. If these shoots formbetween the petioles of the stalk, several petioles have to be strippedoff causing the celery to become smaller and the functional yields to bedecreased.

Vascular Bundle. In celery, the xylem and phloem run vertically throughthe petiole near the epidermis in groups or traces called vascularbundles.

Vacuum. Means the negative pressure (in inches mercury) required torupture or break the celery petiole, measured in inches of mercury.

Vegetable material. Means products that are derived from, but notlimited to, vegetables, fruit, grains and other plants.

DETAILED DESCRIPTION OF THE INVENTION

Celery cultivar ADS-19 is a hollow petiole stem celery (Apium graveolensvar. dulce) resulting from a cross between a stem celery parent (Apiumgraveolens var. dulce) and a celeriac or root celery (Apium graveolensvar. rapaceum). ADS-19 has been selected for numerous generations todevelop a variety that retains the hollow stem without forming a swollenroot while having the appearance of a normal tall celery more similar inappearance to varieties developed for production of sticks (i.e. ADS-11,ADS-17, ADS-15). ADS-19 is upright and fairly compact with a shingledhabit. ADS-19 produces no suckers and has very little heart development.

However, celery cultivar ADS-19 is not as well shingled or refined asADS-15. ADS-19 does have the tendency to have a more open appearance atthe butt because the petioles tend to be larger (deeper) at lower pointon the base of the petiole which possesses a smaller flare. This varietyis particularly important for its very thick and juicy petioles that aremore tender and favorable for consumption. The petioles are also largerin diameter so that they are much more adequate for stuffing with fooditems. ADS-19 is too large for use as a straw and does not have thedurability of ADS-9 or ADS-15.

Celery cultivar ADS-19 has the following morphologic and othercharacteristics (based primarily on data collected at Oxnard, Calif.).

TABLE 1 VARIETY DESCRIPTION INFORMATION Maturity: 87 days in Oxnard,California Plant Height: 87.0 cm Number of Outer Petioles (>40 cm): 9.8Number of Inner Petioles (<40 cm): 2.9 Stalk Shape: Fairly cylindricalStalk Conformation: Open to compact Heart Formation: Sparse PetioleLength (from butt to first joint): 44.2 cm Petiole Length Class: Long(>30 cm) Petiole Width (at midpoint): 1.83 cm Petiole Thickness (atmidpoint): 1.25 cm Wall Thickness at Side: 0.45 cm Cross Section HollowColor (un-blanched at harvest): MUN 5GY 6/8 (green) Anthocyanin: AbsentStringiness: Stringless Ribbing: Smooth Glossiness: Glossy Leaf BladeColor: MUN 5GY 4/6 (dark green) Bolting: Fairly tolerant StressTolerance: Adaxial Crackstem (Boron Deficiency): Tolerant AbaxialCrackstem (Boron Deficiency): Tolerant Leaf Margin Chlorosis (MagnesiumDeficiency): Tolerant Blackheart (Calcium Deficiency): TolerantPithiness (Nutritional Deficiency): Not applicable Feather Leaf:Tolerant Sucker Development: Tolerant Disease Resistance: FusariumYellows, Race 2 (Fusarium oxysporum): Slight tolerance Brown Stem:Tolerant

ADS-19 is similar to celery cultivar ADS-15 but there are severalsignificant differences. ADS-19 has a less compact and refined stalkwhen compared to ADS-15. ADS-19 consistently produces less outerpetioles that are larger (width), and thicker at the side when comparedto ADS-15. While the ADS-19 petioles are considerably thicker, they alsohave a lower percentage dry weight and decreased tolerance to a vacuumand side pressure (see Tables 2-3). This is indicative of a variety thathas a much juicier, fleshier, mouth feel when compared to ADS-15 andespecially ADS-9. ADS-19 is less fibrous and has a more enjoyable eatingfeel. ADS-19 produces a petiole that is much smoother than ADS-15 andADS-9 (Table 2-3). ADS-19 is consistently paler in color in the petioleand often shows a paler hue in the leaf while the Munsell color scaledoes not readily distinguish the difference in the leaf hue (Table -33).

While ADS-19 makes petioles that are valuable for general consumption asa hollow celery that can be stuffed, ADS-19 is not as nice for use as astraw for sipping drinks. While durable compared to most other root orleaf celeries (Tables 4-7) it is not as durable as ADS-9 and ADS-15(Tables 2-3). While durable enough for use as a straw, ADS-19 is alsogenerally too large for sipping. While ADS-15 can be harvested at anearlier maturity (Tables 2-3) in order to get a smaller diameter strawmore ideal for sipping, this is not possible for ADS-19.

This invention is also directed to methods for producing a celery plantby crossing a first parent celery plant with a second parent celeryplant, wherein the first parent celery plant or second parent celeryplant is the celery plant from cultivar ADS-19. Further, both the firstparent celery plant and second parent celery plant may be from cultivarADS-19. Therefore, any methods using celery cultivar ADS-19 are part ofthis invention: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using celery cultivar ADS-19 as atleast one parent are within the scope of this invention.

Additional methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method suchas microprojectile-mediated delivery, DNA injection, electroporation andthe like. More preferably, expression vectors are introduced into planttissues by using either microprojectile-mediated delivery with abiolistic device or by using Agrobacterium-mediated transformation.Transformant plants obtained with the protoplasm of the invention areintended to be within the scope of this invention.

FURTHER EMBODIMENTS OF THE INVENTION

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes”. Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed line.

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed celery plants,using transformation methods as described below to incorporatetransgenes into the genetic material of the celery plant(s).

Expression Vectors for Celery Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent which maybe an antibiotic or a herbicide, or genes that encode an altered targetwhich is insensitive to the inhibitor. A few positive selection methodsare also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene, isolated from transposonTn5, which when placed under the control of plant regulatory signalswhich confers resistance to kanamycin (Fraley et al., Proc. Natl. Acad.Sci. U.S.A., 80:4803 (1983)). Another commonly used selectable markergene is the hygromycin phosphotransferase gene which confers resistanceto the antibiotic hygromycin (Vanden Elzen et al., Plant Mol. Biol.,5:299 (1985)).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferase,the bleomycin resistance determinant (Hayford et al., Plant Physiol.86:1216 (1988), Jones et al., Mol. Gen. Genet, 210:86 (1987), Svab etal., Plant Mol. Biol. 14:197(1990), Hille et al., Plant Mol. Biol. 7:171(1986)). Other selectable marker genes confer resistance to herbicidessuch as glyphosate, glufosinate or bromoxynil (Comai et al., Nature317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618 (1990) andStalker et al., Science 242:419-423 (1988)).

Selectable marker genes for plant transformation that are not ofbacterial origin include, for example, mouse dihydrofolate reductase,plant 5-enolpyruvylshikimate-3-phosphate synthase and plant acetolactatesynthase (Eichholtz et al., Somatic Cell Mol. Genet. 13:67 (1987), Shahet al., Science 233:478 (1986), Charest et al., Plant Cell Rep. 8:643(1990)).

Another class of marker genes for plant transformation require screeningof presumptively transformed plant cells rather than direct geneticselection of transformed cells for resistance to a toxic substance suchas an antibiotic. These genes are particularly useful to quantify orvisualize the spatial pattern of expression of a gene in specifictissues and are frequently referred to as reporter genes because theycan be fused to a gene or gene regulatory sequence for the investigationof gene expression. Commonly used genes for screening presumptivelytransformed cells include

α-glucuronidase (GUS), α-galactosidase, luciferase and chloramphenicol,acetyltransferase (Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987),Teeri et al., EMBO J. 8:343 (1989), Koncz et al., Proc. Natl. Acad. SciU.S.A. 84:131 (1987), DeBlock et al., EMBO J. 3:1681 (1984)).

In vivo methods for visualizing GUS activity that do not requiredestruction of plant tissues are available (Molecular Probes publication2908, IMAGENE GREEN, p. 1-4 (1993) and Naleway et al., J. Cell Biol.115:151a (1991)). However, these in vivo methods for visualizing GUSactivity have not proven useful for recovery of transformed cellsbecause of low sensitivity, high fluorescent backgrounds and limitationsassociated with the use of luciferase genes as selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells (Chalfie et al., Science 263:802 (1994)). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Celery Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred”.Promoters which initiate transcription only in certain tissue arereferred to as “tissue-specific”. A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter which is under environmental control. Examples ofenvironmental conditions that may effect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter which is active under mostenvironmental conditions.

A. Inducible Promoters

An inducible promoter is operably linked to a gene for expression incelery. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence which is operably linkedto a gene for expression in celery. With an inducible promoter the rateof transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system whichresponds to copper (Meft et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize which responds to benzenesulfonamide herbicide safeners(Hershey et al., Mol. Gen Genetics 227:229-237 (1991) and Gatz et al.,Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz etal., Mol. Gen. Genetics 227:229-237 (1991). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression incelery or the constitutive promoter is operably linked to a nucleotidesequence encoding a signal sequence which is operably linked to a genefor expression in celery.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)). The ALS promoter, Xba1/Ncol fragment 5′ to the Brassicanapus ALS3 structural gene (or a nucleotide sequence similarity to saidXba1/Ncol fragment), represents a particularly useful constitutivepromoter. See PCT application WO 96/30530.

C. Tissue-specific or Tissue-preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin celery. Optionally, the tissue-specific promoter is operably linkedto a nucleotide sequence encoding a signal sequence which is operablylinked to a gene for expression in celery. Plants transformed with agene of interest operably linked to a tissue-specific promoter producethe protein product of the transgene exclusively, or preferentially, ina specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene (Murai et al., Science 23:476-482(1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. U.S.A.82:3320-3324 (1985)); a leaf-specific and light-induced promoter such asthat from cab or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985)and Timko et al., Nature 318:579-582 (1985)); an anther-specificpromoter such as that from LAT52 (Twell et al., Mol. Gen. Genetics217:240-245 (1989)); a pollen-specific promoter such as that from Zm13(Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993)) or amicrospore-preferred promoter such as that from apg (Twell et al., Sex.Plant Reprod. 6:217-224 (1993)).

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992), Close, P. S.,Master's Thesis, Iowa State University (1993), Knox, C., et al.,AStructure and Organization of Two Divergent Alpha-Amylase Genes fromBarley”, Plant Mol. Biol. 9:3-17 (1987), Lerner et al., Plant Physiol.91:124-129 (1989), Fontes et al., Plant Cell 3:483-496 (1991), Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991), Gould et al., J. Cell.Biol. 108:1657 (1989), Creissen et al., Plant J. 2:129 (1991), Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984), Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6(1981).

According to a preferred embodiment, the transgenic plant provided forcommercial production of foreign protein is celery. In another preferredembodiment, the biomass of interest is seed. For the relatively smallnumber of transgenic plants that show higher levels of expression, agenetic map can be generated, primarily via conventional RFLP, PCR andSSR analysis, which identifies the approximate chromosomal location ofthe integrated DNA molecule. For exemplary methodologies in this regard,see Glick and Thompson, Methods in Plant Molecular Biology andBiotechnology, CRC Press, Boca Raton 269:284 (1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes That Confer Resistance to Pests or Disease and That Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant line can be transformed with a clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266:789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262:1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78:1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btδ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Va., forexample, under ATCC Accession Nos. 40098, 67136, 31995 and 31998.

C. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

D. A vitamin-binding protein such as avidin. See PCT application US93/06487, the contents of which are hereby incorporated by reference.The application teaches the use of avidin and avidin homologues aslarvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor), Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor 1), Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus α-amylase inhibitor).

F. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor), and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyperaccumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference.

M. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci 89:43 (1993), ofheterologous expression of a cecropin-β, lytic peptide analog to rendertransgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. SeeTaylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

P. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10:1436 (1992). The cloning and characterization of a gene which encodesa bean endopolygalacturonase-inhibiting protein is described by Toubartet al., Plant J. 2:367 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

S. A lettuce mosaic potyvirus (LMV) coat protein gene introduced intoLactuca sativa in order to increase its resistance to LMV infection. SeeDinant et al., Molecular Breeding. 1997, 3: 1, 75-86.

2. Genes That Confer Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988), and Miki et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance conferred by mutant5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus PAT bar genes), and pyridinoxy or phenoxy proprionic acidsand cyclohexones (ACCase inhibitor-encoding genes). See, for example,U.S. Pat. No. 4,940,835 to Shah, et al., which discloses the nucleotidesequence of a form of EPSPS which can confer glyphosate resistance. ADNA molecule encoding a mutant aroA gene can be obtained under ATCCaccession number 39256, and the nucleotide sequence of the mutant geneis disclosed in U.S. Pat. No. 4,769,061 to Comai. See alsoUmaballava-Mobapathie in Transgenic Research. 1999, 8: 1, 33-44 thatdiscloses Lactuca sativa resistant to glufosinate. European patentapplication No. 0 333 033 to Kumada et al., and U.S. Pat. No. 4,975,374to Goodman et al., disclose nucleotide sequences of glutamine synthetasegenes which confer resistance to herbicides such as L-phosphinothricin.The nucleotide sequence of a phosphinothricin-acetyl-transferase gene isprovided in European application No. 0 242 246 to Leemans et al. DeGreefet al., Bio/Technology 7:61 (1989), describe the production oftransgenic plants that express chimeric bar genes coding forphosphinothricin acetyl transferase activity. Exemplary of genesconferring resistance to phenoxy proprionic acids and cyclohexones, suchas sethoxydim and haloxyfop are the Acc1-S1, Acc1-S2 and Acc1-S3 genesdescribed by Marshall et al., Theor. Appl. Genet. 83:435 (1992).

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3:169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

D. Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants. See Hattori et al., Mol. Gen.Genet. 246:419, 1995. Other genes that confer tolerance to herbicidesinclude a gene encoding a chimeric protein of rat cytochrome P4507A1 andyeast NADPH-cytochrome P450 oxidoreductase (Shiota et al., PlantPhysiol., 106:17, 1994), genes for glutathione reductase and superoxidedismutase (Aono et al., Plant Cell Physiol. 36:1687, 1995), and genesfor various phosphotransferases (Datta et al., Plant Mol. Biol. 20:619,1992).

E. Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306; 6,282,837;5,767,373; and international publication WO 01/12825.

F. Modified bolting tolerance in plants for example, by transferring agene encoding for gibberellin 2-oxidase. See U.S. Pat. No. 7,262,340.Bolting has also been modified using non-transformation methods. SeeWittwer, S. H., et al. (1957) Science. 126(3262): 30-31; Booij, R. etal., (1995) Scientia Horticulturae. 63:143-154; and Booij, R. et al.,(1994) Scientia Horticulturae. 58:271-282.

3. Genes That Confer or Contribute to a Value-Added Trait, Such as:

A. Increased iron content of the celery, for example by transforming aplant with a soybean ferritin gene as described in Goto et al., ActaHorticulturae. 2000, 521, 101-109.

B. Decreased nitrate content of leaves, for example by transforming acelery with a gene coding for a nitrate reductase. See for exampleCurtis et al., Plant Cell Report. 1999, 18: 11, 889-896.

C. Increased sweetness of the celery by transferring a gene coding formonellin, that elicits a flavor 100,000 times sweeter than sugar on amolar basis. See Penarrubia et al., Biotechnology. 1992, 10: 561-564.

D. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.USA 89:2625 (1992).

E. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene), Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene), Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus lichenifonnis α-amylase), Elliot et al., PlantMolec. Biol. 21:515 (1993) (nucleotide sequences of tomato invertasegenes), SØgaard et al., J. Biol. Chem. 268:22480 (1993) (site-directedmutagenesis of barley α-amylase gene), and Fisher et al., Plant Physiol.102:1045 (1993) (maize endosperm starch branching enzyme II).

4. Genes that Control Male-Sterility

A. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT. See international publication WO 01/29237.

B. Introduction of various stamen-specific promoters. See internationalpublications WO 92/13956 and WO 92/13957.

C. Introduction of the barnase and the barstar genes. See Paul et al.,Plant Mol. Biol. 19:611-622, 1992).

Methods for Celery Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical, plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, GlickB. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993)pages 89-119.

A. Agrobacterium-mediated Transformation

One method for introducing an expression vector into plants is based onthe natural transformation system of Agrobacterium. See, for example,Horsch et al., Science 227:1229 (1985), Curtis et al., Journal ofExperimental Botany. 1994, 45: 279, 1441-1449, Torres et al., Plant cellTissue and Organic Culture. 1993, 34: 3, 279-285, Dinant et al.,Molecular Breeding. 1997, 3: 1, 75-86. A. tumefaciens and A. rhizogenesare plant pathogenic soil bacteria which genetically transform plantcells. The Ti and Ri plasmids of A. tumefaciens and A. rhizogenes,respectively, carry genes responsible for genetic transformation of theplant. See, for example, Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991).Descriptions of Agrobacterium vector systems and methods forAgrobacterium-mediated gene transfer are provided by Gruber et al.,supra, Miki et al., supra, and Moloney et al., Plant Cell Reports 8:238(1989). See also, U.S. Pat. No. 5,591,616 issued Jan. 7, 1997.

B. Direct Gene Transfer

Several methods of plant transformation, collectively referred to asdirect gene transfer, have been developed as an alternative toAgrobacterium-mediated transformation. A generally applicable method ofplant transformation is microprojectile-mediated transformation whereinDNA is carried on the surface of microprojectiles measuring 1 to 4 μm.The expression vector is introduced into plant tissues with a biolisticdevice that accelerates the microprojectiles to speeds of 300 to 600 m/swhich is sufficient to penetrate plant cell walls and membranes.Russell, D. R., et al. Pl. Cell. Rep. 12(3, January), 165-169 (1993),Aragao, F. J. L., et al. Plant Mol. Biol. 20(2, October), 357-359(1992), Aragao, F. J. L., et al. Pl. Cell. Rep. 12(9, July), 483-490(1993). Aragao, Theor. Appl. Genet. 93: 142-150 (1996), Kim, J.;Minamikawa, T. Plant Science 117: 131-138 (1996), Sanford et al., Part.Sci. Technol. 5:27 (1987), Sanford, J. C., Trends Biotech. 6:299 (1988),Klein et al., Bio/Technology 6:559-563 (1988), Sanford, J. C., PhysiolPlant 7:206 (1990), Klein et al., Biotechnology 10:268 (1992).

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome and spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO J., 4:2731 (1985), Christouet al., Proc Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine have also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985) and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Saker, M.; Kuhne, T. Biologia Plantarum 40(4): 507-514(1997/98), Donn et al., In Abstracts of VIIth International Congress onPlant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990); D'Halluin etal., Plant Cell 4:1495-1505 (1992) and Spencer et al., Plant Mol. Biol.24:51-61 (1994). See also Chupean et al., Biotechnology. 1989, 7: 5,503-508.

Following transformation of celery target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic line. The transgenic line could then be crossed,with another (non-transformed or transformed) line, in order to producea new transgenic celery line. Alternatively, a genetic trait which hasbeen engineered into a particular celery cultivar using the foregoingtransformation techniques could be moved into another line usingtraditional backcrossing techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove an engineered trait from a public, non-elite inbred line into anelite inbred line, or from an inbred line containing a foreign gene inits genome into an inbred line or lines which do not contain that gene.As used herein, “crossing” can refer to a simple X by Y cross, or theprocess of backcrossing, depending on the context.

Single-Gene Conversions

When the term celery plant, cultivar or celery line are used in thecontext of the present invention, this also includes any single geneconversions of that line. The term “single gene converted plant” as usedherein refers to those celery plants which are developed by a plantbreeding technique called backcrossing wherein essentially all of thedesired morphological and physiological characteristics of a cultivarare recovered in addition to the single gene transferred into the linevia the backcrossing technique. Backcrossing methods can be used withthe present invention to improve or introduce a characteristic into theline. The term “backcrossing” as used herein refers to the repeatedcrossing of a hybrid progeny back to one of the parental celery plantsfor that line, backcrossing 1, 2, 3, 4, 5, 6, 7, 8 or more times to therecurrent parent. The parental celery plant which contributes the genefor the desired characteristic is termed the nonrecurrent or donorparent. This terminology refers to the fact that the nonrecurrent parentis used one time in the backcross protocol and therefore does not recur.The parental celery plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent asit is used for several rounds in the backcrossing protocol (Poehlman &Sleper, 1994; Fehr, 1987). In a typical backcross protocol, the originalcultivar of interest (recurrent parent) is crossed to a second line(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until a celeryplant is obtained wherein essentially all of the desired morphologicaland physiological characteristics of the recurrent parent are recoveredin the converted plant, in addition to the single transferred gene fromthe nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalline. To accomplish this, a single gene of the recurrent cultivar ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original line. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross, one ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new line but that can be improvedby backcrossing techniques. Single gene traits may or may not betransgenic, examples of these traits include but are not limited to,male sterility, modified fatty acid metabolism, modified carbohydratemetabolism, herbicide resistance, resistance for bacterial, fungal, orviral disease, insect resistance, enhanced nutritional quality,industrial usage, yield stability and yield enhancement. These genes aregenerally inherited through the nucleus. Several of these single genetraits are described in U.S. Pat. Nos. 5,777,196, 5,948,957 and5,969,212, the disclosures of which are specifically hereby incorporatedby reference.

Tissue Culture

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of celery andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Teng et al., HortScience. 1992, 27: 9,1030-1032 Teng et al., HortScience. 1993, 28: 6, 669-1671, Zhang et al.,Journal of Genetics and Breeding. 1992, 46: 3, 287-290, Webb et al.,Plant Cell Tissue and Organ Culture. 1994, 38: 1, 77-79, Curtis et al.,Journal of Experimental Botany. 1994, 45: 279, 1441-1449, Nagata et al.,Journal for the American Society for Horticultural Science. 2000, 125:6, 669-672, and Ibrahim et al., Plant Cell, Tissue and Organ Culture.(1992), 28(2): 139-145. It is clear from the literature that the stateof the art is such that these methods of obtaining plants are routinelyused and have a very high rate of success. Thus, another aspect of thisinvention is to provide cells which upon growth and differentiationproduce celery plants having the physiological and morphologicalcharacteristics of variety ADS-19.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, meristematic cells, andplant cells that can generate tissue culture that are intact in plantsor parts of plants, such as leaves, pollen, embryos, roots, root tips,anthers, pistils, flowers, seeds, petioles, suckers and the like. Meansfor preparing and maintaining plant tissue culture are well known in theart. By way of example, a tissue culture comprising organs has been usedto produce regenerated plants. U.S. Pat. Nos. 5,959,185; 5,973,234 and5,977,445 describe certain techniques, the disclosures of which areincorporated herein by reference.

Additional Breeding Methods

This invention also is directed to methods for producing a celery plantby crossing a first parent celery plant with a second parent celeryplant wherein the first or second parent celery plant is a celery plantof cultivar ADS-19. Further, both first and second parent celery plantscan come from celery cultivar ADS-19. Thus, any such methods usingcelery cultivar ADS-19 are part of this invention: selfing, backcrosses,hybrid production, crosses to populations, and the like. All plantsproduced using celery cultivar ADS-19 as at least one parent are withinthe scope of this invention, including those developed from cultivarsderived from celery cultivar ADS-19. Advantageously, this celerycultivar could be used in crosses with other, different, celery plantsto produce the first generation (F₁) celery hybrid seeds and plants withsuperior characteristics. The cultivar of the invention can also be usedfor transformation where exogenous genes are introduced and expressed bythe cultivar of the invention. Genetic variants created either throughtraditional breeding methods using celery cultivar ADS-19 or throughtransformation of cultivar ADS-19 by any of a number of protocols knownto those of skill in the art are intended to be within the scope of thisinvention.

The following describes breeding methods that may be used with celerycultivar ADS-19 in the development of further celery plants. One suchembodiment is a method for developing cultivar ADS-19 progeny celeryplants in a celery plant breeding program comprising: obtaining thecelery plant, or a part thereof, of cultivar ADS-19 utilizing said plantor plant part as a source of breeding material, and selecting a celerycultivar ADS-19 progeny plant with molecular markers in common withcultivar ADS-19 and/or with morphological and/or physiologicalcharacteristics selected from the characteristics listed in Table 1.Breeding steps that may be used in the celery plant breeding programinclude pedigree breeding, backcrossing, mutation breeding, andrecurrent selection. In conjunction with these steps, techniques such asRFLP-enhanced selection, genetic marker enhanced selection (for exampleSSR markers) and the making of double haploids may be utilized.

Another method involves producing a population of celery cultivar ADS-19progeny celery plants, comprising crossing cultivar ADS-19 with anothercelery plant, thereby producing a population of celery plants, which, onaverage, derive 50% of their alleles from celery cultivar ADS-19. Aplant of this population may be selected and repeatedly selfed or sibbedwith a celery cultivar resulting from these successive filialgenerations. One embodiment of this invention is the celery cultivarproduced by this method and that has obtained at least 50% of itsalleles from celery cultivar ADS-19.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. For example, see Fehr and Walt, Principles of CultivarDevelopment, p 261-286 (1987). Thus the invention includes celerycultivar ADS-19 progeny celery plants comprising a combination of atleast two cultivar ADS-19 traits selected from the group consisting ofthose listed in Table 1 or the cultivar ADS-19 combination of traitslisted in the Summary of the Invention, so that said progeny celeryplant is not significantly different for said traits than celerycultivar ADS-19 as determined at the 5% significance level when grown inthe same environmental conditions. Using techniques described herein,molecular markers may be used to identify said progeny plant as a celerycultivar ADS-19 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and preferably thetraits are measured on plants grown under the same environmentalconditions. Once such a variety is developed its value is substantialsince it is important to advance the germplasm base as a whole in orderto maintain or improve traits such as yield, disease resistance, pestresistance, and plant performance in extreme environmental conditions.

Progeny of celery cultivar ADS-19 may also be characterized throughtheir filial relationship with celery cultivar ADS-19, as for example,being within a certain number of breeding crosses of celery cultivarADS-19. A breeding cross is a cross made to introduce new genetics intothe progeny, and is distinguished from a cross, such as a self or a sibcross, made to select among existing genetic alleles. The lower thenumber of breeding crosses in the pedigree, the closer the relationshipbetween celery cultivar ADS-19 and its progeny. For example, progenyproduced by the methods described herein may be within 1, 2, 3, 4 or 5breeding crosses of celery cultivar ADS-19.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cell tissue cultures from which celery plants can beregenerated, plant calli, plant clumps and plant cells that are intactin plants or parts of plants, such as leaves, pollen, embryos, roots,root tips, anthers, pistils, flowers, seeds, petioles, suckers and thelike.

Industrial Uses of Celery Cultivar ADS-19

Celery may be used in a variety of manner including but not limited to,use in salads, soups, being filled, injected or stuffed with cheese,soybean, vegetable, peanut butter or dairy type products and serveduncooked, baked like a manicotti pasta, or filled, and cooked in amanner similar to enchiladas, or stuffed, battered and deep fried like ajalapeno popper.

Preparation of the Celery Stick of ADS-19

In the preparation of a product for the marketplace, raw celery isharvested by hand or machine in the field similar to standard stemcelery and placed in bins, totes or cartons and cooled. Cooling willusually be performed by utilizing hydro-vac cooling, hydro cooling orforced aircooling methods typical of most raw vegetables, but othermethods may be used.

The initial processing steps may include cleaning steps often used forraw vegetables to assure cleanliness and food safety.

Once the celery is cooled, the cool process will be maintained withinthe range of 33° F. to 40° F. unless the specific product or processrequires a break in that chain.

In the present invention, a whole celery stalk is trimmed to remove thebutt and foliage. The celery may either be cut by hand or mechanicalmeans such as a saw or knife. It may also be cut with a water knife orsimilar type advancements in cutting technology.

The celery of the present invention is a novel and natural stuffablecelery, is the result of cutting the petioles of hollow celery in amanner that maintains the integrity of the celery. Maintenance of theintegrity of the stem is especially critical because holes or cracks inthe product will result in an unusable product.

The water knife/water jet cutter is a special technology which has beenadopted specially for cutting hollow celery. Due to the hollow nature ofthe product, conventional knives and saws have a greater opportunity tocollapse the tube at the point of impact, thus causing the tube torupture, split or crack.

However, the water knife cuts by a very high pressure stream of waterand air (over 37,000 psi) being passed through a very small orifice ornozzle (approximately, but not limited to, 0.0007 cm in diameter). Thehollow celery passing through this high pressure stream of air and wateris cut with no risk of straw collapse, because no physical pressure isapplied to the hollow celery. See, for example, U.S. Pat. Nos.3,974,725, 4,601,156, 4,753,808, 6,308,600, 4,751,094, and 5,916,354.

Water is first run through a pre-chiller which drops the temperature tobetween 34° and 36° F. The water is then run into an intensifier whereit is run through a filtration system to remove impurities. Theintensifier then compresses water and air independently. A stream ofwater is then injected into the air such that the air acts as thecarrier and the water as the abrasive. This mixture passes through a setof cutting nozzles (>37,000 psi) that have an orifice between 0.0003 cmand 0.0010 cm in diameter.

Several water jet nozzles are placed in series at intervals that matchthe length of the celery straws that are desired. The whole hollowcelery stalk is then passed through these nozzles or knives on aconveyor. As the celery stalk passes though the pressurized wateremanating from each nozzle the celery stalk is cut into a celery stick.The length of the celery stick will depend on the specific product orthe requirements of the consumer.

Additional cleaning steps typical of raw, semi-processed and processedvegetables may be utilized to assure cleanliness and food safety. Thesesteps may include chlorine or other cleansing/sterilizers in rinse waterapplied via a drenching or water bath system. Typical solutions includechlorine and water at a concentration of approximately 750 ppm orcleansing/sterilizers at their suggested use rates. Other technologiesmay be utilized as appropriate or acceptable.

All products should be handled from this step forward in an asepticenvironment following general HACCP procedures in order to ensure foodsafety.

The cut and cleaned hollow celery stick is sorted or graded by size andquality based on standards established for the specific products, usesor customer requirements. The celery will be run through a metaldetector following processing or prior to or following packaging toensure that no metal has contaminated the product.

Hollow celery sticks will be packaged and sealed in various containertypes according to the customer's requirements. The cool process remainsunbroken for this product, as it is sold as a perishable product.

Hollow celery sticks may be handled in a slightly dehydrated (wilted orlimber) state and then rehydrated by the consumer by placing in acontainer with clean water for several minutes. The consumer may trimthe ends with a knife to improve freshness, appearance, or adjust thelength.

Sanitation of Celery

Prior to delivery of the celery to a food injection site, a celery willneed to undergo a sanitation process. The celery may be sanitized by anynumber of methods which include but are not limited to, the following:ascorbic acid and peroxyacetic acid also known as TSUNAMI, sodiumhypochlorite (chlorine), bromine products (sodium hypobromine), chlorinedioxide, ozone based systems, hydrogen peroxide products, trisodiumphosphate, quaternary ammonium products, ultraviolet light systems,solar radiation, nuclear radiation, irradiation, steam, ultra heattreatments, ultra cold treatments and high pressure pasteurization. See,for example, U.S. Pat. Nos. 7,220,381, 5,945,146 and 4,753,808 and USPublication No. 2004/0191382, Zagory, D. 1999. Liao, C., Cooke, P. H.2001. Response to Trisodium Phosphate Treatment of Salmonella ChesterAttached to Fresh-Cut Green Pepper Slices. Canadian J. Micro. 47:25-32;Jongen, W., ed. 2005. Improving the Safety of Fresh Fruit andVegetables. C.H.I.P.S., Weimar, Tex.

Another method that could be used to sanitize and wash the celery stickswould be the use of cool or warm air that is blown through the hollowcelery stick, the hollow celery is then placed in a centrifuge or shakerto remove as much moisture as possible, the straws are then allowed tonaturally drain by standing on end. The hollow celery sticks are thenplaced in a vacuum to remove all additional moisture and finally placedin a desiccator.

Once the celery has been sanitized, raw or cooked hollow celery stickmay be stuffed with an injection system specifically modified to matchthe diameter of the hollow celery stick and the consistency of the foodproduct being injected.

Possible methods of delivering the hollow celery sticks to the foodinjection location for injecting various food products into the hollowcelery sticks include but are not limited to mechanical means includinghydraulic, pneumatic and electrical. The equipment that is used forthese methods includes but is not limited to belt conveyors, flat,flighted or grooved, made of vinyl or rubber. Shakers or vibratingmachines for up and down and/or side to side motions or fast back andforth motions could also be used. Step-motion orientators that create aback and forth, side to side or up and down motion or elevatororientators that create vertical or any degree of incline or declinecould be used as well as transfer slides that create any degree ofincline or decline. Finally, transfer belts, including flat, flightedand grooved made of plastice or rubber could be used to deliver thecelery to the food product injection location.

Delivery of the hollow celery sticks and the food product to theinjection location may also take place manually and include equipmentsuch as containers, tables, transfer jigs, placement jigs, andsemi-automatic feeders.

Once the hollow celery sticks and the food product have been deliveredto the injection site the method of injection may include but is notlimited to: hydraulic, pneumatic, electrical or water injection. Theequipment that may be used in this process includes but is not limitedto injection needles and injection tubes. The force required to injectthe food product into the hollow celery stick could be created throughforce air or vacuum pressure based on either positive or negativepressure. The force required to inject the food product into the hollowcelery sticks could also be created through forced or vacuum waterpressure under either positive or negative pressure.

The injection of the food product into the hollow celery sticks couldalso be done manually. The equipment that may be necessary for thisprocess includes but is not limited to injections needles, injectiontubes, plastics or rubber basters, pastry bags or frosting bags,frosting tips, and semi-automatic injectors.

Depending on the particular food product being injected, another coolingprocedure may be required to re-establish an appropriate temperature.This cooling procedure, if required, may take place prior or justfollowing packaging in customer specified packaging. The cold processmust be maintained throughout shipment and delivery to the customer.

Each type of cooked, stuffed product may have an entirely differenttreatment or preparation process.

Once the celery is sanitized and injected with the appropriate foodproduct, they are packaged according to length and may be packaged inany number of methods according to the specifications of the customer.The product of the present invention may be packaged in numerous typesof packages including but not limited to rigid plastic, flexible film,solid fiber, poly sleeves, plastic sleeves, poly bags, plastic bags,natural decomposable bags, natural decomposable sleeves, packages thatmay be opened and sealed, rigid containers like clam shells, packageswith different permeability properties, packages with built-in vents,packages with specialized pores or any combination thereof, or anycombination thereof. Variations in the packaging may include differentgas exchange rates which may occur due to different permeability ortransmission properties of the package materials themselves or due tovents or specialized pores built into the packaging. See for example,U.S. Pat. Nos. 4,753,808, and 4,586,313.

TABLES

In the tables that follow, the traits and characteristics of celerycultivar ADS-19 are given compared to other check cultivars.

Table 2 shows a comparison between ADS-15, ADS-19 and ADS-9 in a trialgrown in Oxnard, Calif. The trial was transplanted Mar. 6, 2007 at apopulation of 58,080 plants per acre. Production conditions were typicalwithout significant stress.

TABLE 2 ADS-15 ADS-19 ADS-9 Apium Apium Apium graveolens graveolensgraveolens L. var L. var L. var dulce dulce dulce Maturity (days) 84 8787 Mean Plant Height 87.5 87.3 85.9 (cm) Mean Whole Plant 0.728 0.7980.513 weight (kg) Mean Trim Plant 0.605 0.739 0.494 weight (kg) MeanNumber of 0 0 15.6 Suckers Mean Joint 47.6 44.2 50.1 Length(cm) MeanNumber of 10.6 10.4 14.2 Outer Petioles >40 cm Mean Number of 4 4 4.7Inner Petioles <40 cm Mean Width of Outer 15.7 17.9 10.4 Petioles @midrib (mm) Mean Depth of Outer 12 12.3 7.1 Petioles @ midrib (mm) MeanWall Thickness 3.7 4.5 2 at Sides of Petiole (mm) Mean Number of 7 14.913.1 22 inch Straws per Plant Mean Straw Yield per 0.264 0.295 0.233Plant (kg) Mean Weight per 17.7 22.5 10.6 Straw (g) Number of 7 inch865,392 760,848 1,277,760 Straws per Acre Straw Yield per Acre 15,33317,134 13,533 (kg)

As shown in Table 2, compared to ADS-15, ADS-19 is slightly shorter tothe joint, produces slightly less petioles, and weighs approximately 10%more in the whole stalk. ADS-19's main points of differentiation are awider petiole (1 4%) that is 22% thicker and gives the finished strawyield per plant and acre 12% more weight in spite of yielding 14% lessstraws than ADS-15. This wider, thicker petiole in ADS-19 is moreconducive and desirable for stuffing while the ADS-15 is preferred forstraws for drinking beverages.

Table 3 shows a comparison between ADS-15, ADS-19 and ADS-9 in a trialgrown in Oxnard, Calif. The trial was transplanted Mar. 13, 2007 at apopulation of 58,080 plants per acre. Production conditions were typicalwith out significant stress.

TABLE 3 ADS-15 Apium ADS-19 ADS-9 graveolens Apium Apium L. vargraveolens graveolens dulce L. var dulce L. var dulce Maturity (days) 8087 87 Mean Plant Height (cm) 91.8 87 83 Mean Whole Plant weight 0.7310.801 0.486 (kg) Mean Trim Plant weight 0.599 0.712 0.453 (kg) MeanNumber of Suckers 0 0 18.2 Mean Joint Length(cm) 46.2 44.2 47.8 MeanNumber of Outer 10.3 9.8 13.3 Petioles >40 cm Mean Number of Inner 3.12.9 4.6 Petioles <40 cm Mean Width of Outer 16.2 18.3 11.1 Petioles @midrib (mm) Mean Depth of Outer 12.0 12.5 7.0 Petioles @ midrib (mm)Mean Wall Thickness at 3.7 4.6 2.6 Sides of Petiole (mm) Mean Number of7 inch 12.9 11.7 20.2 Straws per Plant Mean Straw Yield per Plant 0.2550.265 0.206 (kg) Mean Weight per Straw (g) 19.8 22.6 10.2 Number of 7inch Straws 749,232 679,536 1,173,216 per Acre Straw Yield per Acre (kg)14,810 15,391 11,964

As shown in Table 3, compared to ADS-15, ADS-19 is slightly shorter tothe joint, produces slightly less petioles and weighs approximately 10%more in the whole stalk. ADS-19's main points of differentiation are awider petiole (13%) that is 25% thicker and gives the finished strawyield per plant and acre 4 % more weight in spite of yielding 10% lessstraws than ADS-15. This wider thicker petiole in ADS-19 is moreconducive and desirable for stuffing while the ADS-15 is preferred forstraws for drinking beverages.

Table 4 shows a comparison between ADS-19 and two other hollow stemcelery (ADS-9 and ADS-15) Blanco de Veneto (Apium graveolens L. varrapaceum) a celeriac or root celery and Afina (Apium graveolens L. varsecalinum) a leaf celery in a trial grown in Oxnard, Calif. The trialwas transplanted December 2007 at a population of 58,080 plants peracre. Production was during a period when bolting pressure was severe.Harvest occurred Apr. 21, 2008 at 115 days maturity.

TABLE 4 ADS-15 ADS-9 ADS-19 Blanco de Veneto Afina Apium Apium ApiumApium Apium graveolens L. var graveolens L. var graveolens L. vargraveolens L. var graveolens L. var dulce dulce dulce rapaceum secalinumMean Plant Height (cm) 95.1 96.7 100.6 73.2 80.7 Mean Whole Plant weight0.865 0.814 0.911 0.497 0.744 (kg) Mean Root Diam. (cm) 0.0 0.0 0.0 51.80.0 Mean Root weight (kg) 0.0 0.0 0.0 0.072 0.0 Mean Joint Length (cm)49.3 57.2 52.6 29.7 45.4 Mean Number of Outer 8.3 11.9 8.1 6.8 22.6Petioles >40 cm Mean Number of Inner 5.1 8.9 5.0 7.3 85.2 Petioles <40cm Mean Number of Suckers 0.0 17.0 0.0 2.0 101.0 Mean Seed Stem Length8.8 45.7 9.9 35.9 23.3 (cm) Mean Width of Outer 17.7 11.7 18.4 7.3 6.0Petioles @ midrib (mm) Mean Depth of Outer 16.6 11.3 16.9 6.9 5.2Petioles @ midrib (mm) Mean Vacuum (in/Hg) 22.3 25.9 21.1 10.1 11.5 MeanWall Thickness at 4.2 2.8 4.5 2.2 1.4 Sides of Petiole (mm) Mean WallThickness at 1.2 1.0 1.5 0.6 0.6 Inside of Petiole Cup (mm) MeanPressure Required 2257.2 1198.0 1886.8 817.5 505.8 to Rupture Side WallMean Pressure Required 343.4 398.7 325.3 149.4 164.3 to Rupture Wall @Inside of Petiole Cup Leaf Color (Muncell) 5gy 4/6 5gy 4/8 5gy 4/6 5gy4/4 5gy 4/6  Petiole Color (Muncell) 5gy 6/6 5gy 6/6 5gy 6/8 5gy 5/6 5gy5/10 Petiole Smoothness slight rib slight rib smooth ribbed ribbed

As shown in Table 4, ADS-15 and ADS-19 are the most similar in theseresults and have a significantly thicker and stronger petiole side walland petiole cup than Blanco de Veneto (Apium graveolens L. var rapaceum)a celeriac or root celery and Afina (Apium graveolens L. var secalinum)a leaf celery. While ADS-15 is designed for use as a straw ADS-19 is alarger, less fibrous celery line designed for stuffing and use foractual consumption as opposed to a straw. The ADS-19 and ADS-15varieties (Apium graveolens L. var dulce) are contrasted to Blanco deVeneto (Apium graveolens L. var rapaceum) a celeriac or root celery andAfina (Apium graveolens L. var secalinum) a leaf celery. The Blanco deVeneto measurements for root diameter and depth are included but do notrepresent a fully mature root; however the petioles are at full size andrepresent a reasonable comparison to ADS-15 and ADS-19. ADS-15 andADS-19 are fairly bolting tolerant and significantly more boltingtolerant than the other celeriac and leaf celery varieties. ADS-15 has asignificantly wider and deeper petiole, thicker side wall and inside ofcup wall, requires significantly more pressure to rupture the side wallof the petiole and inside of cup of the petiole when compared to ADS-9and Blanco de Veneto and Afina. The ability to withstand a vacuum wasalso significantly improved when compared to Blanco de Veneto and Afina,however the ability to withstand vacuum pressure by ADS-19 was less thanADS-9. ADS-15 was not as thick at the petiole side wall and inside ofthe cup as ADS-19 but still significantly thicker than Blanco de Veneto(Apium graveolens L. var rapaceum) a celeriac or root celery and Afina(Apium graveolens L. var secalinum) a leaf celery. ADS-15 and ADS-19have no suckers when compared with ADS-9 and Afina. Afina which isessentially a stalk comprised primarily of suckers is very typical ofleaf celery while ADS-9 still has only a few suckers characteristic ofits leaf celery progenitor and is more similar to a stem celery with ahigher sucker count. This contrasts to ADS-15 and ADS-19 which have aceleriac as one of the originating parents. While celeriacs may vary inthe number of suckers possessed, dependent on the variety, ADS-15 andADS-19 are more similar to tall stem varieties like ADS-11, ADS-17 andADS-18 which typically have no suckers.

Table 5 shows a comparison between ADS-19, Afina a hollow-stem leafcelery and numerous hollow-stem celeriacs, commercial andrepresentatives from the United States germplasm collection. Allcomparisons were generated from a trial harvested May 29, 2008 inOxnard, Calif. The trial was transplanted Feb. 27, 2008 at a populationof 58,080 plants per acre. Production was during a period whenconditions were fairly normal and free from most stresses. This datadoes show that there may have been marginal bolting pressure, butpressure was light and the presence of seed stems as an indication thatthe varieties were particularly sensitive. Harvest occurred May 29, 2008at 92 days maturity.

TABLE 5 Blanco de ADS-19 Veneto Monarch Afina PI 193454 PI 179171 ApiumApium Apium Apium Apium Apium graveolens L. graveolens L. graveolens L.graveolens L. graveolens L. graveolens L. var dulce var rapaceum varrapaceum var secalinum var rapaceum var rapaceum Mean Plant Height (cm)91.4 49.3 45.9 55.6 53.9 67.9 Mean Plant Width (cm) 34.8 36.1 27.7 41.644.4 45.0 Mean Whole Plant weight 0.967 0.368 0.301 0.531 0.409 0.465(kg) Mean Trimmed Plant 0.901 0.3295 0.1935 0.198 0.295 0.249 Weight(kg) Mean Number of Suckers 0.0 0.0 0.0 103.8 0.0 25.8 Mean RootDiameter (cm) 0.0 56.8 61.9 0.0 82.0 66.0 Mean Root Depth (cm) 0.0 64.276.1 0.0 96.0 80.0 Mean Root weight (kg) 0.0 0.114 0.115 0.0 0.258 0.213Mean Joint Length (cm) 44.6 20.0 17.8 24.8 27.8 37.6 Mean Number ofOuter 10.0 9.7 12.1 7.4 12.6 11.9 Petioles (>40 cm) Mean Number of Inner6.6 9.8 6.5 9.2 7.1 5.0 Petioles (<40 cm) Mean Seed Stem Length 0.0 0.00.0 0.0 0.5 1.2 (cm) Mean Width of Outer 18.1 7.3 7.1 6 7.5 7.5 Petioles@ midrib (mm) Mean Depth of Outer 11.9 6.5 6.5 3.9 4.6 6.6 Petioles @midrib (mm) Mean Number of 7 inch 12.4 1.1 0 7 7.6 11.7 Straws per PlantMean Straw Yield per Plant 0.320 0.012 0 0.044 0.061 0.084 (kg) MeanWeight per Straw (g) 25.8 10.5 0 6.3 8 7.2 Number 7″ Straws per Acre720,192 63,888 0 406,560 441,408 679,536 Straw Yield per Acre (kg)18,586 668 0 2,556 3,543 4,879 Mean Vacuum (in/Hg) 25.3 10.7 11.2 11.512.9 14.0 Mean Wall Thickness at 4.18 1.82 2.07 1.26 1.01 1.42 Sides ofPetiole (mm) Mean Wall Thickness at 1.44 0.75 0.55 0.39 0.54 0.37 Insideof Petiole Cup (mm) Mean Pressure Required to 2105 1554 1630 1055 990698 Rupture Side Wall (g) Mean Pressure Required to 661 417 444 414 292145 Rupture Wall @ Inside Of Petiole Cup (g) Mean Percentage Dry 6.2%8.0% 7.7% 8.5% 8.2% 9.2% Weight

As shown in Table 5, ADS-19 is 10% to 21% taller than the hollow-stemleaf celery and celeriac lines. ADS-19 does not have suckers. ADS-19also has a significantly thicker petiole side wall and petiole cup thanthe Apium graveolens L. var secalinum and Apium graveolens L. varrapaceum varieties. While Afina is a typical representative of the leafcelery class which is essentially all suckers, the celeriac's have afairly wide range of suckers from 0 to 50 per stalk. None of theceleriac varieties have the preponderance of suckers as represented byAfina. A significant difference between ADS-19 and the root celery lines(Apium graveolens L. var rapaceum) is the absence of a swollen root(tuber). While the maturity was not sufficient to allow for maximumswelling/yield there was obvious and measureable swelling in theceleriacs. Measurements were taken for width, depth and weight. Noswelling had occurred in ADS-19 stem celery (Apium graveolens L. vardulce) or leaf celery (Apium graveolens L. var secalinum). Differencesbetween ADS-19 when compared with all other classes became especiallypronounced when the petiole width and thickness was measured at themid-rib. The total number of straws or 7 inch petiole segments variedamong all of the lines but when all characteristics, including widththickness, vacuum, rupture pressure and wall thickness were consideredonly ADS-19 made reasonably usable straws. When comparing wallthickness, vacuum and rupture pressure ADS-19 was much more durable thanthe celeriac and leaf celery lines.

Table 6 shows a comparison between ADS-19 and five hollow-stem celeriacsor root celery lines ), that are commercially available or availablefrom the United States germplasm collection. All comparisons weregenerated from a trial harvested May 29, 2008 in Oxnard, Calif. Thetrial was transplanted Feb. 27, 2008 at a population of 58,080 plantsper acre. Production was during a period when conditions were fairlynormal and free from most stresses. This data does show that there mayhave been marginal bolting pressure, but pressure was light and thepresence of seed stems an indication that the varieties wereparticularly sensitive. Harvest occurred May 29, 2008 at 92 daysmaturity.

TABLE 6 ADS-19 PI 261810* PI 164944 PI 169001 PI 176417 PI 178834 ApiumApium Apium Apium Apium Apium graveolens L. graveolens L. graveolens L.graveolens L. graveolens L. graveolens L. var dulce var rapaceum varrapaceum var rapaceum var rapaceum var rapaceum Mean Plant Height (cm)91.4 49.5 81.0 61.2 63.3 81.0 Mean Plant Width (cm) 34.8 42.6 49.5 52.752.8 52.2 Mean Whole Plant weight 0.967 0.426 0.647 0.569 0.474 0.724(kg) Mean Trimmed Plant 0.901 0.308 0.266 0.398 0.274 0.43 Weight (kg)Mean Number of 0.0 3.5 49.6 18.3 35.0 40.0 Suckers Mean Root Diameter0.0 90.0 70.0 70.0 48.3 47.2 (cm) Mean Root Depth (cm) 0.0 100.0 80.072.0 74.0 78.0 Mean Root weight (kg) 0.0 0.375 0.235 0.212 0.073 0.23Mean Joint Length (cm) 44.6 24.0 40.6 31.0 36.1 47.2 Mean Number ofOuter 10.0 6.1 12.8 16.5 13.6 14.9 Petioles (>40 cm) Mean Number ofInner 6.6 17.3 8.1 6.4 5.0 4.8 Petioles (<40 cm) Mean Seed Stem Length0.0 0.0 47.7 4.4 4.0 1.9 (cm) Mean Width of Outer 18.1 7.7 7.6 7.7 6.77.3 Petioles @ midrib (mm) Mean Depth of Outer 11.9 3.5 5.6 5.5 4.7 5.8Petioles @ midrib (mm) Mean Number of 7 inch 12.4 6.5 16.3 12.9 13.420.1 Straws per Plant Mean Straw Yield per 0.320 0.054 0.103 0.103 0.0940.115 Plant (kg) Mean Weight per Straw 25.8 8.3 6.3 8 7 8.2 (g) Number7″ Straws per 720,192 377,520 946,704 749,232 778,272 1,167,408 AcreStraw Yield per Acre (kg) 18,586 3,136 5,982 5,982 5,460 6,679 MeanVacuum (in/Hg) 25.3 12.7 18.1 15.3 16.9 10.6 Mean Wall Thickness at 4.181.21 1.47 1.56 1.5 1.6 Sides of Petiole (mm) Mean Wall Thickness at 1.440.41 0.44 0.66 0.51 0.54 Inside of Petiole Cup (mm) Mean PressureRequired 2105 657 1205 1062 1370 1296 to Rupture Side Wall (g) MeanPressure Required 661 123 375 486 467 415 to Rupture Wall @ Inside OfPetiole Cup (g) Mean Percentage Dry 6.2% 9.2% 9.4% 9.0% 9.7% 8.7% Weight

As shown in table 6, ADS-19 is 10 to 21% taller than all other lines.ADS-19 does not have suckers. The celeriac's represent a fairly widerange of suckers from 0 to 50 per stalk. A significant differencebetween ADS-19 and the root celery lines (rapaceum) is the absence of aswollen root (tuber). While the maturity was not sufficient to allow formaximum swelling/yield there was obvious and measureable swelling.Measurements were taken for width, depth and weight. No swelling hadoccurred in ADS-19. While bolting pressure was very light, Pl 164944 wasobviously very susceptible to bolting. Differences between the hollowpetiole dulce types when compared with all other classes becameespecially pronounced when the petiole width and thickness was measuredat the mid-rib. The total number of straws or 7 inch segments variedamong all of the lines but when all characteristics, including widththickness, vacuum, rupture pressure and wall thickness were consideredonly ADS-19 made reasonably usable straws. When comparing wallthickness, vacuum and rupture pressure ADS-19 was much more durable thanall other lines tested.

Table 7 shows a comparison between ADS-19 and two other hollow petiolestem celeries (ADS-9 and ADS-15), two root celeries, Monarch and Blancode Veneto (Apium graveolens L. var rapaceum) and Afina a leaf celery(Apium graveolens L. var secalinum) in a trial grown in Salinas, Calif.The trial was transplanted Apr. 22, 2007 at a population of 63,000plants per acre. Production was under normal conditions with nostresses. Harvest occurred Jul. 23, 2007 at 92 days maturity.

TABLE 7 ADS-15 ADS-9 ADS-19 Afina Monarch Blanco De Veneto Apium ApiumApium Apium Apium Apium graveolens L. graveolens L. graveolens L.graveolens L. graveolens L. graveolens L. var dulce var dulce var dulcevar secalinum var rapaceum var rapaceum Mean Width of 17.9 12.3 21.1 5.56.5 6 Outer Petioles @ midrib (mm) Mean Depth of 14 9.9 16.4 5 6 6 OuterPetioles @ midrib (mm) Mean Vacuum 15.9 17.4 12.4 8.4 9.1 10.2 (in/Hg)Mean Wall 3.5 3.0 4.8 1.2 1.3 1.3 Thickness at Sides of Petiole (mm)Mean Wall 1.5 1.0 1.5 0.7 0.7 0.7 Thickness at Inside of Petiole Cup(mm) Mean Pressure 1921 2050 2166 1521 975 873 Required to Rupture SideWall (grams) Mean Pressure 454 700 434 201 195 150 Required to RuptureWall @ Inside of Petiole Cup (grams) Mean Percent dry 7.6% 8.6% 7.4%9.1% 9.1% 10.2% weight

As can be seen in Table 7 the petiole width of ADS-19, ADS-15 and ADS-9(Apium graveolens L. var dulce) are significantly greater than thepetiole width of the leaf celery (Apium graveolens L. var secalinum) androot celery (Apium graveolens L. var rapaceum) varieties. The petiole ofADS-9 is approximately 100% larger while the petioles of ADS-15 andADS-19 are at least 175% wider than Afina, Monarch and Blanco de Veneto(Apium graveolens L. var secalinum and Apium graveolens L. varrapaceum). Similarly the depth of the outter petioles at the midrib aresignificantly larger than the Apium varieties with ADS-9 (65% deeper)and ADS-15 and ADS-19 (at least 133%) deeper than Afina, Monarch andBlanco de Veneto (Apium graveolens L. var secalinum and Apium graveolensL. var rapaceum). In combination with thicker side walls, 130% to 269%thicker, a greater capacity to withstand a vacuum, 22% to 71 % moreresilient and greater pressure required to rupture the walls the ADS-19,ADS-15 and ADS-9 varieties are more suited for use as a straw. WhenADS-15 and ADS-19 are compared directly, ADS-19 is found to be moreappropriate for use as a consumed or edible product to be stuffed withedible material, while ADS-15 is more appropriate for use as a straw.ADS-19 has petioles that are 18% wider and 17% deeper than ADS-15 makinga larger hollow tube more suitable for stuffing. The petioles of ADS-19are also 71 % thicker and have a lower percentage dry weight thanADS-15. This lower percentage dry weight correlates with ADS-19 beingjuicier, less fibrous and much more edible. ADS-15 on the other hand ismore suited to a straw with slightly smaller diameter.

Table 8 shows a comparison between ADS-15 and ADS-19 in a trial grown inOxnard, Calif. for the purpose of evaluating the varieties for toleranceto Fusarium oxysporum f. sp. apii race 2. The trial was transplantedAug. 13, 2008 at a population of 50,000 plants per acre. This trial wassown in a research plot that has been specially developed with elevatedFusarium levels.

TABLE 8 ADS-15 ADS-19 Apium graveolens Apium graveolens L. var dulce L.var dulce Mean Plant Height (cm) 107.6 102.6 Mean Whole Plant weight(kg) 0.956 0.764 Mean Trim Plant weight (kg) 0.729 0.589 Mean Number ofSuckers 0 0 Mean Joint Length(cm) 47.9 50.2 Mean Number of Outer 11.38.9 Petioles >40 cm Mean Number of Inner 1.8 2.1 Petioles <40 cm MeanWidth of Outer Petioles 15.1 15.8 @ midrib (mm) Mean Depth of OuterPetioles 13.1 13.0 @ midrib (mm) Mean Wall Thickness at Sides 2.9 3.4 ofPetiole (mm) Mean Number of 7 inch 17.4 14.5 Straws per Plant Mean StrawYield per Plant 0.319 0.338 (kg) Mean Weight per Straw (g) 0.018 0.023Number of 7 inch Straws per 870,000 725,000 Acre Straw Yield per Acre(kg) 15,950 16,900 Mean General Fusarium 4 3.5 Rating (0 = death to 5 =resistant) Mean Fusarium Injury in the 4 3.0 Root (0 = death to 5 =resistant)

As can be seen in Table 8, a comparison of ADS-19 as it performed underconditions with no Fusarium (Tables 2 and 3) to these conditions whereFusarium was severe indicate that there was little impact on the size,yield, length and number of the straws generated. While there was someFusarium present as evidenced by the general Fusarium and root Fusariumratings, however the economic impact was not significant. The dataindicate that celery cultivar ADS-15 has better tolerance to Fusariumthan celery cultivar ADS-19.

DEPOSIT INFORMATION

A deposit of A. Duda & Sons, Inc. proprietary Celery Cultivar ADS-19disclosed above and recited in the appended claims has been made withthe American Type Culture Collection (ATCC), 10801 University Boulevard,Manassas, Va. 20110. The date of deposit was Jun. 23, 2010. The depositof 2,500 seeds was taken from the same deposit maintained by A. Duda &Sons, Inc. since prior to the filing date of this application. Allrestrictions will be removed upon granting of a patent, and the depositis intended to meet all of the requirements of 37 C.F.R. §§1.801-1.809.The ATCC Accession Number is PTA-11090. The deposit will be maintainedin the depository for a period of thirty years, or five years after thelast request, or for the enforceable life of the patent, whichever islonger, and will be replaced as necessary during that period.

1. A seed of celery cultivar ADS-19, representative sample seed of saidcultivar was deposited under ATCC Accession No. PTA-11090.
 2. A celeryplant, or a part thereof, produced by growing the seed of claim
 1. 3. Atissue culture produced from protoplasts or cells from the plant ofclaim 2, wherein said cells or protoplasts are produced from a plantpart selected from the group consisting of leaf, callus, pollen, ovule,embryo, cotyledon, hypocotyl, meristematic cell, root, root tip, pistil,anther, flower, seed, shoot, stem, petiole and sucker.
 4. A celery plantregenerated from the tissue culture of claim 3, wherein the plant hasall of the morphological and physiological characteristics of cultivarADS-19.
 5. A method for producing a celery seed comprising crossing twocelery plants and harvesting the resultant celery seed, wherein at leastone celery plant is the celery plant of claim
 2. 6. A celery seedproduced by the method of claim
 5. 7. A celery plant, or a part thereof,produced by growing said seed of claim
 6. 8. The method of claim 5,wherein at least one of said celery plants is transgenic.
 9. A method ofproducing a herbicide resistant celery plant, wherein said methodcomprises introducing a gene conferring herbicide resistance into theplant of claim
 2. 10. A herbicide resistant celery plant produced by themethod of claim 9, wherein the gene confers resistance to a herbicideselected from the group consisting of glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine, andbenzonitrile.
 11. A method of producing a pest or insect resistantcelery plant, wherein said method comprises introducing a geneconferring pest or insect resistance into the celery plant of claim 2.12. A pest or insect resistant celery plant produced by the method ofclaim
 11. 13. The celery plant of claim 12, wherein the gene encodes aBacillus thuringiensis (Bt) endotoxin.
 14. A method of producing adisease resistant celery plant, wherein said method comprisesintroducing a gene into the celery plant of claim
 2. 15. A diseaseresistant celery plant produced by the method of claim
 14. 16. A methodfor producing a male sterile celery plant, wherein said method comprisestransforming the celery plant of claim 2, with a nucleic acid moleculethat confers male sterility.
 17. A male sterile celery plant produced bythe method of claim
 16. 18. A method of introducing a desired trait intocelery cultivar ADS-19 wherein the method comprises: (a) crossing aADS-19 plant, wherein a representative sample of seed was depositedunder ATCC Accession No. PTA-11090, with a plant of another celerycultivar that comprises a desired trait to produce progeny plantswherein the desired trait is selected from the group consisting ofimproved nutritional quality, industrial usage, male sterility,herbicide resistance, insect resistance, modified seed yield, modifiedlodging resistance, modified iron-deficiency chlorosis and resistance tobacterial disease, fungal disease or viral disease; (b) selecting one ormore progeny plants that have the desired trait to produce selectedprogeny plants; (c) crossing the selected progeny plants with the ADS-19plants to produce backcross progeny plants; (d) selecting for backcrossprogeny plants that have the desired trait and all of the physiologicaland morphological characteristics of celery cultivar ADS-19 listed inTable 1; and (e) repeating steps (c) and (d) two or more times insuccession to produce selected third or higher backcross progeny plantsthat comprise the desired trait and all of the physiological andmorphological characteristics of celery cultivar ADS-19 listed inTable
 1. 19. A celery plant produced by the method of claim 18, whereinthe plant has the desired trait.
 20. The celery plant of claim 19,wherein the desired trait is herbicide resistance and the resistance isconferred to a herbicide selected from the group consisting ofimidazolinone, dicamba, cyclohexanedione, sulfonylurea, glyphosate,glufosinate, phenoxy proprionic acid, L-phosphinothricin, triazine andbenzonitrile.
 21. The celery plant of claim 19, wherein the desiredtrait is insect resistance and the insect resistance is conferred by agene encoding a Bacillus thuringiensis endotoxin.
 22. A method forproducing a celery stick, comprising the steps of a. cutting the celeryplant of claim 2 to remove leaves; b. cutting said celery to removeentire attachment to butt of celery; c. removing heart of said celery;d. cutting said celery into sticks between 1.0 inches and 12.0 inches;and e. sanitizing said celery to produce a celery stick.
 23. A celerystick made by the method of claim 22, wherein the celery stick is cutand filled with consumable products.